Purification and characterization of mRNA cap-binding protein from Drosophila melanogaster embryos

@article{Maroto1989PurificationAC,
  title={Purification and characterization of mRNA cap-binding protein from Drosophila melanogaster embryos},
  author={Federico Garc{\'i}a Maroto and Jos{\'e} Manuel Sierra},
  journal={Molecular and Cellular Biology},
  year={1989},
  volume={9},
  pages={2181 - 2190}
}
A protein with specific affinity for the mRNA cap structure was purified both from the postribosomal supernatant and from the ribosomal high-salt wash of Drosophila melanogaster embryos by m7GTP-Sepharose chromatography. This protein had an apparent molecular mass of 35 kilodaltons (kDa) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a size very different from those of the cap-binding proteins that have been characterized thus far. Drosophila 35-kDa cap-binding protein (CBP… 
Alternatively Spliced Transcripts from the Drosophila eIF4E Gene Produce Two Different Cap-binding Proteins*
TLDR
Two different protein isoforms result from three alternatively spliced transcripts from a single eIF4E gene, which maps to region 67A8-B2 on polytene chromosomes, and vary greatly in the lengths of their 5′-UTRs, suggesting the possibility of complex translational control of expression of the two eif4E isoforms.
Two functionally redundant isoforms of Drosophila melanogaster eukaryotic initiation factor 4B are involved in cap-dependent translation, cell survival, and proliferation.
TLDR
RNA interference experiments suggest that eIF4B is required for cell survival, although only a modest reduction in rate of protein synthesis is observed, and suggests that cap-dependent over IRES-dependent translation initiation in a Drosophila cell-free translation system.
Localization, structure and expression of the gene for translation initiation factor eIF-4E from Drosophila melanogaster
TLDR
Several sequences which may be involved in the regulation of transcription initiation of the eIF-4E gene, except for a consensus TATA box, were found upstream of the putative transcription initiation sites.
embryos Drosophila mRNAs in heat shock protein 70 and reaper Internal ribosome entry site drives cap-independent translation of
TLDR
In vitro translation experiments using wild-type and eIF4E mutant embryonic extracts show that reporter mRNA bearing reaper 5 untranslated region (UTR) is effectively translated in a cap-independent manner.
m7GpppG cap dependence for efficient translation of Drosophila 70-kDa heat-shock-protein (Hsp70) mRNA.
TLDR
Complementary experiments in which eIF-4 was inactivated in vitro using either m7GTP cap analogue or foot-and-mouth-disease virus L protease expression indicated that the cap-dependent translation pathway is required for optimal Hsp mRNA translation.
Proteomic analysis of reaper 5' untranslated region‐interacting factors isolated by tobramycin affinity‐selection reveals a role for La antigen in reaper mRNA translation
TLDR
The data provide evidence of the involvement of La antigen in the translation of rpr and set a protocol for purification of tagged‐RNA‐protein complexes from cytoplasmic extracts.
Internal ribosome entry site drives cap-independent translation of reaper and heat shock protein 70 mRNAs in Drosophila embryos.
TLDR
In vitro translation experiments using wild-type and eIF4E mutant embryonic extracts show that reporter mRNA bearing reaper 5' untranslated region (UTR) is effectively translated in a cap-independent manner.
Distinct domains of Me31B interact with different eIF4E isoforms in the male germ line of Drosophila melanogaster
TLDR
The observations suggest that Me31B might recognize different eIF4E isoforms in different tissues, which could be the key to silencing specific messengers and provide further evidence that alternative forms of eif4E and their interactions with various partners add complexity to the control of gene expression in eukaryotes.
...
...

References

SHOWING 1-10 OF 47 REFERENCES
Protein synthesis in Drosophila melanogaster embryos. Purification and characterization of polypeptide chain-initiation factor 2.
TLDR
The properties of Drosophila elF-2 suggest that this factor may be susceptible to regulation by a mechanism like that operating on rabbit reticulocyte elF2, and data supporting the notion that this affinity is increased in the presence of Mg2+, which impairs the GDP/GTP exchange on elF, are presented.
Immunological detection of the messenger RNA cap-binding protein.
Eukaryotic mRNA cap binding protein: purification by affinity chromatography on sepharose-coupled m7GDP.
A 24,000-dalton polypeptide that binds strongly and can be specifically crosslinked to the 5'-terminal cap structure m7GpppN in eukaryotic mRNAs has been detected in protein synthesis initiation
Amino acid sequence of the mRNA cap-binding protein from human tissues.
TLDR
The amino acid sequence of CBP showed homology to the cap-binding PB2 protein of influenza virus, and the DNA sequence obtained from recombinant lambda phage inserts was found to code for all but one peptide.
Purification of the messenger RNA cap-binding protein using a new affinity medium.
TLDR
The p-aminophenyl gamma-ester of 7-methylguanosine 5'-triphosphate was synthesized and coupled to Sepharose 4B, and the eluted material was enriched nearly 200-fold in the ability to specifically bind 32P-labeled capped oligonucleotides.
Involvement of the 24-kDa cap-binding protein in regulation of protein synthesis in mitosis.
mRNA cap-binding protein: cloning of the gene encoding protein synthesis initiation factor eIF-4E from Saccharomyces cerevisiae
TLDR
Gene disruption experiments showed that the eIF-4E gene is essential for growth and encodes a protein of 213 amino acid residues.
The cap-binding protein complex in uninfected and poliovirus-infected HeLa cells.
...
...